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WO2008148016A1 - Inhibition des signes du vieillissement par l'inhibition de l'activation du facteur nf-kappa b - Google Patents

Inhibition des signes du vieillissement par l'inhibition de l'activation du facteur nf-kappa b Download PDF

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WO2008148016A1
WO2008148016A1 PCT/US2008/064735 US2008064735W WO2008148016A1 WO 2008148016 A1 WO2008148016 A1 WO 2008148016A1 US 2008064735 W US2008064735 W US 2008064735W WO 2008148016 A1 WO2008148016 A1 WO 2008148016A1
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activation
inhibitor
mice
aging
erccl
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WO2008148016A9 (fr
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Paul D. Robbins
Laura J. Niedernhofer
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University of Pittsburgh
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A61Q19/08—Anti-ageing preparations
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/4418—Non condensed pyridines; Hydrogenated derivatives thereof having a carbocyclic group directly attached to the heterocyclic ring, e.g. cyproheptadine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/49—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds
    • A61K8/4906—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with one nitrogen as the only hetero atom
    • A61K8/4926—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with one nitrogen as the only hetero atom having six membered rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
    • A61K2800/74—Biological properties of particular ingredients
    • A61K2800/78—Enzyme modulators, e.g. Enzyme agonists
    • A61K2800/782—Enzyme inhibitors; Enzyme antagonists

Definitions

  • the present invention relates to methods for reducing and/or delaying one or more signs of aging which comprise inhibiting NF-kappa B activation, preferably by blocking the interaction between NF- ⁇ B essential modulator ("NEMO") with IKB kinase- ⁇ (IKK- ⁇ ) at the NEMO binding domain (NBD).
  • NEMO NF- ⁇ B essential modulator
  • IKK- ⁇ IKB kinase- ⁇
  • the disposable soma theory of aging posits that aging is the consequence of accumulation of stochastic molecular and cellular damage (Kirkwood, 2005b).
  • the precise nature of the damage that is responsible for aging- related degenerative changes remains ill-defined, but may include mitochondrial damage, telomere attrition, nuclear dysmorphology, accumulation of genetic mutations, DNA, protein or membrane damage.
  • DNA damage is one type of molecular damage that contributes to aging.
  • the majority of human progerias or syndromes of accelerated aging are caused by inherited mutations in genes required for genome maintenance, including Werner syndrome, Cockayne syndrome, trichothiodystrophy and ataxia telangiectasia (Hasty et al., 2003).
  • both DNA lesions Hamilton et al., 2001
  • genetic mutations caused by DNA damage Dolle et al., 2002
  • mice harboring germ-line mutations that confer resistance to genotoxic stress are long-lived (Maier et al., 2004; Migliaccio et al, 1999).
  • ERCCl-XPF is a highly conserved structure-specific endonuclease that is required for at least two DNA repair mechanisms in mammalian cells: nucleotide excision repair (Sijbers et al., 1996) and DNA interstrand crosslink repair (Niedernhofer et al., 2004). Genetic deletion of either Erccl or Xpfm ' the mouse causes an identical and very severe phenotype (Mc Whir et al., 1993; Tian et al., 2004; Weeda et al., 1997).
  • null mice Embryonic development of null mice is normal, but postnatally they develop numerous symptoms associated with advanced age including epidermal atrophy and hyperpigmentation, visual impairment, cerebral atrophy with cognitive deficits, cerebellar degeneration, hypertension, renal insufficiency, decreased liver function, anemia and bone marrow degeneration, osteopenia, sarcopenia, cachexia, and decreased lifespan (Niedernhofer et al., 2006; Prasher et al., 2005; Weeda et al., 1997, and see International Patent Application Publication No. WO2006/052136).
  • the transcriptome from the liver of Erccl-/- mice was compared to that of old wild type (wt) mice and a highly significant correlation was identified (Niedernhofer et al., 2006). Similar expression changes were also identified in young wt mice after chronic exposure to a DNA damaging agent. This provides direct experimental evidence that DNA damage induces changes that mimic aging at the fundamental level of gene expression.
  • Erccl-/- and old mice share not only broad changes in gene expression, but also endocrine, metabolic and cell signaling changes. This implies that ERCCl- deficient mice are an accurate and rapid model system for studying systemic aging in mammals.
  • ERCCl-XPF deficiency A case of human progeria caused by ERCCl-XPF deficiency with symptoms near-identical to those observed in ERCCl -deficient mice has been reported (Niedernhofer et al., 2006). Therefore function of ERCCl-XPF is conserved from man to mouse and the discovery of what is driving aging-like degenerative changes in ERCCl -deficient mice will have direct implications for human health.
  • NF-KB is a highly conserved, ubiquitously expressed family of transcription factors.
  • NF- ⁇ B is a key regulator of cell survival and proliferation in response to a numerous types of stress including oxidative, genotoxic, mitogenic and inflammatory (Hu and Hung, 2005).
  • NF- ⁇ B family members include p65, c-Rel, ReI- B, p50 and p52.
  • NF- ⁇ B exists as a dimer, with the most common being p50-p65.
  • IKB proteins bind to NF- ⁇ B and mask their nuclear localization signals.
  • IKB kinase IKB kinase
  • NF- ⁇ B Activation of NF- ⁇ B is implicated in numerous pathophysiologic conditions including chronic inflammatory diseases (rheumatoid arthritis, asthma and inflammatory bowel disease) (Tak and Firestein, 2001) and cancer (Pikarsky et al., 2004).
  • NF- ⁇ B is also activated in a variety of tissues of aged rodents relative to young animals, including the skin, liver, kidney, cerebellum, cardiac muscle and gastric mucosa (Bregegere et al., 2006; Giardina and Hubbard, 2002; Helenius et al., 1996a; Helenius et al., 1996b; Korhonen et al., 1997; Xiao and Majumdar, 2000).
  • NF- ⁇ B activation has been implicated in driving cellular senescence (Gosselin and Abbadie, 2003).
  • NF- ⁇ B inhibitors modulating molecular effects associated with aging (Kim et al., 2006; Go et al, 2005).
  • One strategy for inhibiting NF- ⁇ B activation involves inhibiting phosphorylation of IKB proteins, thereby preventing mobilization of NF- ⁇ B into the nucleus.
  • IKK IkB kinase
  • NEMO NF- ⁇ B essential modulator
  • IKKalpha and beta two catalytic subunits
  • the interaction of these subunits allows for phosphorylation of IKK, resulting in activation of kinase activity and subsequent phosphorylation IKB, the cytoplasmic inhibitor of NF-kB.
  • IkB is subsequently degraded which releases NF- ⁇ B and reveals a nuclear localization signal, allowing NF- ⁇ B to shuttle to the nucleus and activate pro- inflammatory genes.
  • a short peptide (TALDWSWLQTE; SEQ ID NO: 1) derived from the
  • NBD Nemo Binding Domain of IKK ⁇ blocks association of NEMO with IKKbeta, preventing NF- ⁇ B activation (di Meglio et al., 2005; May et al., 2000). Importantly, the NBD peptide does not affect basal activity of NF- ⁇ B, which is required for mouse development and tissue homeostasis (Li et al., 1999; Nenci et al., 2007; Rudolph et al., 2000; Tanaka et al., 1999), but only suppresses induction of NF- ⁇ B activity in response to inflammatory stimuli and certain types of stress (March et al., 2000).
  • NBD peptide Systemic administration of NBD peptide is not toxic to mice and inhibition of NF- ⁇ B by this peptide correlates with therapeutic response in numerous models of chronic inflammation (Dai et al., 2004; Dasgupta et al., 2004; Dave et al., 2006; di Meglio et al., 2005; Jimi et al., 2004).
  • the use of peptides to block this interaction is disclosed in U.S. Patent Nos. 6,864,355 and 7,049,395 and United States Patent Application Publication No. 2006/0293244.
  • Compound A Another inhibitor of NF- ⁇ B that has been identified is a small molecule inhibitor of IKK- ⁇ , referred to as "Compound A,” which is a 2- amino-3-cyna-4-alkyl-6-(2-hydroxyphenyl) pyridine derivative.
  • the present invention relates to methods for reducing and/or delaying one or more signs of aging which comprise inhibiting NF- ⁇ B activation. It is based, at least in part, on the discovery that a peptide which inhibits IKK- ⁇ interaction with NEMO, linked to a transducing peptide, inhibits the development of various indicia of senescence in a murine model of aging, Erccl "/ ⁇ mice.
  • compositions comprising a NF -KB activation inhibitor which is a NBD peptide comprising a polylysine transduction peptide.
  • a NF -KB activation inhibitor which is a NBD peptide comprising a polylysine transduction peptide.
  • such compositions may be used according to the methods of the invention to reduce and/or delay one or more signs of aging.
  • FIGURE 1 Schematic diagram of mErccl targeting constructs.
  • exon 7 is interrupted with a neo cassette.
  • hypomorphic allele ( ⁇ ) a small deletion was placed at the very C-terminus of the protein to mimic human ERCCl and a neo cassette was placed in intron 9.
  • J, knock-down allele
  • exon 7 was fused with the cDNA of exon 8-10 and the entire region floxed with loxP sites recognized by Cre-recombinase.
  • the neomycin cassette situated in the 3'UTR disrupts Erccl expression, if not removed by Cre recombinase.
  • FIGURE 2 Immunodetection of XPF in protein extracts isolated from the liver of Erccl mutant mice. The genotype of the mice is indicated above the lanes. Below the blots are indicated the calculated level of XPF expression in the Erccl mutant mice, relative to wild type (100%).
  • FIGURE S Clonogenic survival assay measuring the sensitivity of
  • Wild-type data is represented by a circle; Erccl ' ⁇ by a square; Erccl '/A by a triangle, and ErccT 1' by an open square.
  • FIGURE 4 Lifespan of mice expressing various levels of ERCCl- XPF compared to wt C57B1/6 mice (100%; adapted from (Rowlatt et al, 1976)). Wild-ype data is represented by a diamond; ErccT ⁇ by a square; and Erccl '/A by a triangle.
  • FIGURE 5 Up-regulation of NF- ⁇ B in aged wt and progeroid Erccl '/A mouse liver relative to young wt mice. Sections of cryopreserved liver were stained with Dapi to identify nuclei (blue), actin to highlight the cytoplasm (red) and for the p65 subunit of NF- ⁇ B (green), [this figure is in black and white]
  • FIGURE 6 Exposure paradigm for pilot study to determine the impact of 8K-NBD on quality of life in progeroid Erccl ' ⁇ mice. 8K-NBD was dissolved in 5% DMSO in PBS and injected intraperitoneally. A littermate Erccl '/A mouse was treated with an equal volume of vehicle only (PBS). The study was concluded at 19 wks when the mouse treated with vehicle only became profoundly ataxic and cachectic, requiring euthanization.
  • FIGURE 7A-B Images of sibling Erccl '/A mice, one chronically treated with 8K-NBD, the other vehicle only.
  • dystonia dotted arrow
  • kyphosis a mice treated with 8K-NBD.
  • a mouse systemically treated with 8K-NBD maintained body weight significantly longer than untreated mice (circles) or a mouse treated with vehicle only (triangles).
  • FIGURE 10 Exposure paradigm for pilot study to determine the impact of 8K-NBD on histopathologic changes in progeroid Erccl '/& mice. 8K-NBD was dissolved in 5% DMSO in PBS and injected i.p. FIGURE I lA-B. A. Paraffin-embedded liver sections of liver.
  • FIGURE 12 Treatment schedule for third series of experiments; treatment with 8K-NBD or negative control 8k-mNBD.
  • FIGURE 13 Table showing phenotypic changes associated with aging in Erccl '/A mice treated with 8K-NBD or negative control 8K-mNBD.
  • FIGURE 14A-D Comparison of levels of neurodegeneration, as determined by glial fibrillary acidic protein (GFAP) staining, in (A) a young wild- type mouse; (B) a 2-year old (aged) wild-type mouse; (C) a Erccl '/A mouse treated with 8K-NBD according to the regimen of FIGURE 12; and (D) a Erccl '/A mouse treated with negative control 8K-mNBD.
  • GFAP glial fibrillary acidic protein
  • FIGURE 15A-D Comparison of levels of hepatocellular senescence, as measured by staining for pi 6 protein, in (A) a young wild-type mouse; (B) a 2-year old (aged) wild-type mouse; (C) a Erccl '/A mouse treated with 8K-NBD according to the regimen of FIGURE 12; and (D) a Erccl '/A mouse treated with negative control 8K-mNBD.
  • FIGURE 16A-F Comparison of osteoporotic changes in Erccl '/A mice treated with 8K-NBD or negative control 8K-mNBD. Gross macroscopic and cross- sectional views from micro-CT are shown.
  • A gross macroscopic view of vertebra of wild-type, untreated 17 week old mouse;
  • B cross-sectional view of vertebra of wild- type untreated mouse shown in (A);
  • C gross macroscopic view of vertebra of sibling Erccl ⁇ /A mouse treated with 8K-mNBD;
  • D cross-sectional view of vertebra of Erccl ⁇ /A mouse of (C);
  • E gross macroscopic view of vertebra of sibling Erccl '/A mouse treated with 8K-NBD;
  • F cross-sectional view of vertebra oiErccl '/A mouse of (E).
  • FIGURE 17 Family tree to create genetic depletion of NF- ⁇ B through crossing a p65 + " Ercc + ' parent with a Ercc +/” parent and producing p65 + " Ercc " " f 1 progeny.
  • FIGURE 18 Survival of Ercc " ⁇ mice (diamonds) compared with a p65 +/" Ercc "7" mice (squares).
  • FIGURE 19A-D Fluorescently labeled protein transduction domains
  • the PTD is 6-His (His-His-His-His-His-His (SEQ ID NO: 3)) ;
  • the PTD is 6-R (Arg-Arg-Arg-Arg-Arg-Arg (SEQ ID NO:4)) ;
  • the PTD is 8K (Lys-Lys-Lys-Lys-Lys-Lys-Lys-Lys-Lys-Lys-Lys (SEQ ID NO:5));
  • the PTD is Con-P (Ala-Arg-Phe-Leu-Glu-His-Gly-Ser-Asp-Lys-Ala-Thr (SEQ ID NO:6)).
  • FIGURE 20 Transduction of mouse skin by 8K.
  • FIGURE 21 Transduction of mouse skin by peptides with or without cream.
  • NF- ⁇ B activation means the process whereby NF- ⁇ B is freed from its cytoplasmic complex, enters the nucleus, binds its cognate regulatory elements and enhances gene transcription. Important aspects of NF- ⁇ B activation include (i) binding of the regulatory subunit named "NF- ⁇ B essential modulator” (NEMO or IKK ⁇ ) to IKK ⁇ and IKKalpha, activating the IKK complex (ii) phosphorylation of IKB, (iii) transport of NF-kB through the nuclear membrane, (iv) DNA binding of NF- ⁇ B, and (v) transactivation by NF- ⁇ B.
  • An inhibitor of NF- ⁇ B activation may inhibit activation of any of these five aspects.
  • Inhibitors of NEMO binding include peptides comprising a NEMO binding domain "NBD", referred to herein as "NBD peptides".
  • NBD peptides which may further comprise a protein transduction domain, include peptides disclosed in United States Patent No. 6,864,355, United States Patent No. 7,049,395, and/or United States Patent Application Publication No. US 2006/0293244 (each of which is incorporated by reference herein in its entirety).
  • transduction domain-containing peptides which may be comprised in NBD peptides for use according to the invention include transduction domain-containing peptides disclosed in United States Patent No. 6,881,825, International Patent Application No. PCT/US03/04632, United States Patent
  • the transduction domain-containing peptide is a polylysine of between four and twelve lysine residues, preferably eight lysine residues.
  • Other examinples of transduction domain-containing peptides are 6-His, 6-R, and Con-P.
  • the NBD peptide comprises (i) a polylysine transduction peptide of between four and twelve lysine residues, preferably eight lysine residues; and (ii) a NEMO binding domain with a sequence which is either (a) TALD WSWLQTE (SEQ ID NO:1) or (b) a sequence which is at least 90 percent homologous to SEQ ID NO: 1 or (c) a 9-11 amino acid sequence which differs from SEQ ID NO: 1 in no more than two amino acids or (d) a 8-11 amino acid sequence which differs from SEQ ID NO:1 in no more than three amino acids.
  • the NBD peptide may be between about 12 and 50 amino acids in length, or between about 12 and 30 amino acids in length, or between about 12 and 20 amino acids in length. Amino acid sequences which neither function in transduction or NEMO binding may, without limitation, be functionally essentially silent or may, for example, improve the stability of the peptide.
  • the NBD peptide may be KKKKKKKK-GG-TALDWSWLQTE (SEQ ID NO:2); said peptide may be comprised in a pharmaceutical composition, optionally further comprising a suitable pharmaceutical carrier.
  • the inhibitor of NF- ⁇ B activation may be a 2-amino-3-cyna-4-alkyl-6-(2-hydroxyphenyl) pyridine derivative such as, but not limited to, Compound A (Murata et al., 2003; Mustafa and Leverve, 2001; Ziegelbauer et al., 2005).
  • the inhibitor of NF- ⁇ B activation may be BAYl 1-7082 or BAYl 1-7085 (Axxora L.L.C., San Diego, CA) (Petegnief et al., 2001, Neuroscience 104:223).
  • the inhibitor of NF- ⁇ B activation may be SC-514 (Kishore et al., 2003, J Biol. Chem./ 278(35):32861.
  • the inhibitor of NF- ⁇ B activation may be MGl 32 (Calbiochem, La Jolla, CA).
  • the inhibitor of NF- ⁇ B activation may be tosyl-Phe-chloromethylketone ("TPCK").
  • the inhibitor of NF- ⁇ B activation may be (N-6-chloro-7-methoxy-9H- ⁇ -carbolin-8-yl)-2-methylnicotinamide (ML120B, Wen et al., 2006, J. Pharm. Exp. Ther. 317:989-1001.
  • the inhibition of NF-KB activation may be Celastrol (Lee, et al., 2006, Biochem. Pharmacol. 72(10): 1311- 1321).
  • the inhibition of NF-KB activation may be PG201 (Shin, et al., 2005, Biochem. Biophys. Res. Common. 331(4): 1469-1477).
  • theinhibition of NF-KB activation may be MLN0415 (Millenium Pharmaceuticals, Cambridge, MA).
  • the present invention may be used to inhibit the development or progression of one or more signs of aging, including, but not limited to, epidermal atrophy, epidermal hyperpigmentation, rhytid (wrinkles), photoaging of the skin, hearing loss, visual impairment, cerebral atrophy, cognitive deficits, trembling, ataxia, cerebellar degeneration, hypertension, renal insufficiency, renal acidosis, incontinence, decreased liver function, hypoalbuminemia, hepatic accumulation of glycogen and triglycerides, anemia, bone marrow degeneration, osteopenia, kyphosis, degenerative joint disease, intervertebral disc degeneration, sarcopenia, muscle weakness, dystonia, increased peroxisome biogenesis, increased apoptosis, decreased cellular proliferation, cachexia, and decreased lifespan.
  • signs of aging including, but not limited to, epidermal atrophy, epidermal hyperpigmentation, rhytid (wrinkles), photoaging of the skin
  • “Inhibiting the development" of a sign of aging means delaying the onset, slowing the progression, or reducing the manifestation, of a sign of aging.
  • the present invention may be used to improve age-related performance in a geriatric subject.
  • “Improving performance” refers to any aspect of performance, including cognitive performance or physical performance, such as, but not limited to, the ability to be self-sufficient, to take care of (some but not necessarily all) personal needs, to be ambulatory or otherwise mobile, or interaction with others.
  • the present invention may be used to prolong survival of a geriatric subject, for example, relative to an age-matched, clinically comparable control not treated according to the invention.
  • the present invention provides for a method of inhibiting one or more signs of aging in a subject in need of such treatment, comprising administering, to the subject, an effective amount of an inhibitor of NF- ⁇ B activation.
  • the present invention provides for a method of improving age-related performance in a geriatric subject, comprising administering to a subject an effective amount of an inhibitor of NF- ⁇ B activation.
  • the present invention provides for a method of prolonging survival of a geriatric subject, comprising administering, to the s subject, an effective amount of an inhibitor of NF- ⁇ B activation.
  • the inhibitor of NF- ⁇ B activation may be administered systemically to achieve distribution throughout the body or may be administered to achieve a local effect.
  • the route of administration may be selected depending on the intended effect.
  • systemic administration, to achieve therapeutic levels throughout the body may be achieved using an inhibitor suitable for distribution throughout the body, administered via any standard route, including but not limited to oral, intravenous, inhalation, subcutaneous, or intramuscular routes.
  • Non-limiting examples of local administration include, but are not limited to, intrathecal administration to treat central nervous system manifestations of aging, ocular instillation to treat visual disturbances, intramuscular injection may be used to treat muscle wasting, topical administration to prevent or reverse skin aging etc.
  • Topical formulations may include administering the NF- ⁇ B activation inhibitor, optionally comprised in microsphere, microcapsule, or liposome, in a cream, lotion, organic solvent, or aqueous solution.
  • Inhibitors according to the invention may be administered in a suitable pharmaceutical carrier ⁇ e.g. sterile water, normal saline, phosphate buffered saline, etc.).
  • a suitable pharmaceutical carrier e.g. sterile water, normal saline, phosphate buffered saline, etc.
  • inhibitors may be administered as a solution, as a suspension, in solid form, in a sustained release formulation, in a topical cream formulation, etc.
  • an inhibitor may be incorporated into a microcapsule, nanoparticle or liposome.
  • An effective dose may be calculated by determining the amount needed to be administered to produce a concentration sufficient to achieve the desired effect in the tissue to be treated, taking into account, for example, route of administration, bioavailability, half-life, and the concentration which achieves the desired effect in vitro or in an animal model system, using techniques known in the art.
  • Non-limiting examples of doses of NBD peptide inhibitors include between 0.1 and 50 mg/kg, or between 1 and 25 mg/kg, or between 2 and 20 mg/kg, or about 2 mg/kg, or about 10 mg/kg, which may be administered daily, at least 5 times a week, at least 3 times a week, at least twice a week, at least once a week, at least twice a month, at least once a month, at least once every three months, or at least once every six months.
  • a subject may be treated with a NBD peptide inhibitor using a regimen comprising a loading period followed by a maintenance period, wherein the loading period includes treatment, with 1-20 mg/kg or 2-10 mg/kg, daily or every other day for a period of 5-10 days, followed by a maintenance period which includes 1-10 mg/kg, or 10-50 mg/kg, given once a week, twice a week, three times a week, every other week, or once a month.
  • the dose may be between about 0.1 and 20 mg/kg, or between about 0.3 and 10 mg/kg, or between about 2 and 8 mg/kg, or about 5 mg/kg; where Compound B is the inhibitor, the dose may be between about 0.1 and 40 mg/kg, or between about 0.3 and 20 mg/kg.
  • the dose may be between about 0.1 and 10 mg/kg or between about 1 and 10 mg/kg or about 5 mg/kg; where SC-514 is the inhibitor, the dose may be such that the resulting local concentration at the site of treatment is up to 100 ⁇ M for 100% inhibition of NF- KB, as the IC 50 is 11.2 ⁇ M or 8-20 ⁇ M (Kishori 2003); where MGl 32 is the inhibitor, the dose may be such that the resulting local concentration at the site of treatment is between about 0.1 and 0.5 ⁇ M; where TPCK is the inhibitor, the dose may be such that the resulting local concentration at the site of treatment is between about 5-10 ⁇ M; where ML 120B is the inhibitor, the dose may be such that the resulting local concentration at the site of treatment is up to 20 ⁇ M (Wen 2006), as the IC 50 is 60 nM; where Celastrol is the inhibitor, the dose may be between about 1 and 20 mg/kg or between about 10 mg/
  • genomic sequence of Erccl exon 7 was fused to a cDNA encoding exons 8-10 and the fusion floxed with loxP sites.
  • a neomycin cassette was inserted in the 3'UTR.
  • XPF the essential binding partner of ERCCl
  • mice primary mouse embryonic fibroblasts isolated from both mouse strains are proportionately more sensitive to DNA damaging agents than wt cells, but less sensitive than ERCCl -null cells (FIGURE 3). Therefore by titrating the endogenous level of ERCCl-XPF, we have four mouse strains (wt, Erccl '/A , ErccV ⁇ , and Erccl-/-) with different capacities for DNA repair. The maximum lifespan of each mouse strain was measured and compared to wt C57B1/6 mice (FIGURE 4). ErccT 1' mice were found to have, at maximum, a lifespan of 1 month. Erccl ' ⁇ mice had a maximum lifespan of 7 months.
  • Erccl ' ⁇ * ⁇ mice had a normal maximum lifespan. However their median lifespan was significantly reduced compared to wt mice [90 vs. 130 wks, respectively (Rowlatt et al., 1976)]. This demonstrates that capacity for molecular repair, in particular DNA repair, is a direct determinant of lifespan.
  • Erccl ⁇ /& mice develop the same premature aging symptoms as Erccl-/- mice (Niedernhofer et al., 2006). However the age at onset of the symptoms is delayed; Erccl "/ ⁇ mice are asymptomatic for the first 8 wks of life. Beginning at 8 wks, the mice begin to display a constellation of progressive symptoms associated with advanced age including: trembling, ataxia, cerebral atrophy, renal acidosis, decreased liver function, hypoalbuminemia, bone marrow degeneration, osteopenia, kyphosis, dystonia, muscle wasting, growth retardation, cachexia and loss of vision.
  • mice The phenotype of the mice is remarkably homogenous with all mice displaying an identical spectrum of symptoms in a highly predictable order of appearance. This indicates that the Erccl " ⁇ strain is a uniquely accurate and rapid model system for studying mammalian aging and many of the debilitating symptoms associated with human aging.
  • NF- ⁇ B transcription factor is activated in a variety of tissues of aged rodents relative to young animals including the liver, kidney, cerebellum, skin, cardiac muscle and gastric mucosa (Bregegere et al., 2006; Giardina and Hubbard, 2002; Helenius et al., 1996a; Helenius et al., 1996b; Korhonen et al., 1997; Xiao and Majumdar, 2000).
  • liver levels of the p65 subunit of NF- ⁇ B are elevated (Helenius et al., 2001).
  • Activation of NF- ⁇ B has been implicated in driving cellular senescence (Gosselin and Abbadie, 2003).
  • this transcription factor may play a direct role in driving aging and its associated degenerative processes.
  • Hepatocytes from Erccl "7" and Erccl ⁇ /A mice show numerous characteristics of senescence including large polyploid nuclei, intranuclear inclusions and intracellular lipid deposition (Niedernhofer et al., 2006).
  • mice treated with 8K-NBD had improved ambulation and general appearance and maintained weight longer (FIGURE 8).
  • Motor strength and coordination were measured weekly in the mice by testing their ability to cross a balance beam (FIGURE 9A) and to remain suspended from a wire (FIGURE 9B).
  • mice display large polyploid nuclei (FIGURE 1 IA, 10X) characteristic of senescent hepatocytes (Gupta, 2000).
  • hepatocytes from a mouse treated with 8K-NBD showed a much higher cytoplasm to nuclear ratio.
  • hepatic architecture was effaced in the liver of the 8K-NBD treated mouse, suggestive of liver regeneration (Michalopoulos and DeFrances, 2005). SKM of
  • FIG. 12 a different treatment regimen was used (FIGURE 12) in which treatment with 8K-NBD or a negative control peptide, 8K- mNBD (having a mutated sequence, KKKKKKKKGGTALDASALQTE (SEQ ID NO:7)) was initiated earlier than the above regimens at approximately 5 weeks (8K- NBD or 8K-mNBD, 10 mg/kg) administered intraperitoneally three times per week).
  • 8K- NBD or 8K-mNBD 10 mg/kg administered intraperitoneally three times per week.
  • FIGURE 13 presents a table showing the phenotypic changes associated with aging in mice treated with 8k-NBD or negative control 8k-mNBD.
  • the overall aging score was improved by 30 percent in 8K-NBD versus 8K-mNBD-treated mice, indicating a significant delay in the onset of age-related symptoms corresponding to over a decade in human life.
  • FIGURE 14A-D shows a comparison of levels of neurodegeneration, as determined by glial fibrillary acidic protein (GFAP) staining, in young and senescent (aged, 2 yr) wild-type mice as well as Erccl '/A mice treated with either 8K- NBD or negative control 8K-mNBD according to the regimen of FIGURE 12 at the age of 18 weeks.
  • the GFAP staining pattern of the ErccV /& mouse treated with 8K- NBD more closely resembles that of the young wild-type mouse relative to the 8K- mNBD-treated animal.
  • FIGURE 15A-D presents a comparison of levels of hepatocellular senescence, as measured by staining for pi 6 protein, in young and senescent wild- type mice as well as ErccV ⁇ mice treated with either 8K-NBD or negative control 8K-mNBD according to the regimen of FIGURE 12 at the age of 18 weeks.
  • the staining pattern of the 8K-mNBD treated mouse more closely resembles the pattern in senescent wild-type hepatocytes, whereas the 8K-NBD-treated animal's staining pattern more closely resembles that of a young, healthy mouse.
  • FIGURE 16A-F shows a comparison of osteoporotic changes in ErccV / ⁇ mice treated with 8K-NBD or negative control 8K-mNBD and a wild-type littermate at 17 weeks of age. The degree of osteoporotic changes appears greatest in the 8K-mNBD-treated animal.
  • NF- ⁇ B is activated in ErccV /& mice, a mouse model that based on prior precedent (Niedernhofer et al., 2006) ages rapidly as a consequence of defective DNA repair.
  • EXAMPLE GENETIC DEPLETION OF NF- ⁇ B Mice genetically depleted in NF- ⁇ B were produced by crossing a p65 + " Ercc +/" parent with a Ercc +I ⁇ parent and producing p65 +/" Ercc ' ' ' f 1 progeny (FIGURE 17). When the survival of these mice was studied, it was found that the median, although not the maximum, life span was increased in the p65 +/" Ercc '1' mice (FIGURE 18). This supports the conclusion that reduction in the level of NF- ⁇ B activity improves the quality of life, delaying the onset of age-related degenerative changes that limit life in individual members of the species, without significantly extending the lifespan of that species. 10.
  • Chipchase M.D., O'Neill, M. and Melton, D. W. (2003) Characterization of premature liver polyploidy in DNA repair (Erccl)-deficient mice. Hepatology, 38, 958-966.
  • Betaine suppresses ' proinflammatory signaling during aging: the involvement of nuclear factor- ⁇ B via nuclear factor-inducing kinase/I ⁇ B kinase and mitogen-activated protein kinases. J. Gerontol, 6OA, 1252-1264.
  • Migliaccio E., Giorgio, M., MeIe, S., Pelicci, G., Reboldi, P., Pandolfi, P.P., Lanfrancone, L. and Pelicci, P. G. (1999)
  • the p66shc adaptor protein controls oxidative stress response and life span in mammals. Nature, 402, 309-313.
  • Epithelial NEMO links innate immunity to chronic intestinal inflammation. Nature, 446, 557-561.

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Abstract

La présente invention concerne des procédés et compositions visant à limiter et/ou à retarder l'apparition d'un ou plusieurs signes de vieillissement en inhibant l'activation du facteur NF-kappa B. L'invention se fonde, au moins en partie, sur la découverte selon laquelle un peptide inhibant l'interaction entre la sous-unité IKK-ß et la protéine NEMO, peptide qui est relié à un peptide transducteur, inhibe l'apparition de divers signes de sénescence chez un modèle murin du vieillissement, les souris Ercc1-1?.
PCT/US2008/064735 2007-05-25 2008-05-23 Inhibition des signes du vieillissement par l'inhibition de l'activation du facteur nf-kappa b Ceased WO2008148016A1 (fr)

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EP2319488A3 (fr) * 2009-10-02 2013-06-12 Johnson & Johnson Consumer Companies, Inc. Compositions comprenant un inhibiteur de NFkB et un promoteur de la tropoélastine
EP2572701A3 (fr) * 2010-10-25 2013-10-16 Johnson & Johnson Consumer Companies Inc. Compositions comprenant un rétinoïde et inhibiteur de NFkB et leurs procédés d'utilisation
RU2501552C2 (ru) * 2010-08-31 2013-12-20 Федеральное государственное бюджетное учреждение науки Институт биологии Коми научного центра Уральского отделения Российской академии наук Средство для увеличения продолжительности жизни и способ его применения
US9370474B2 (en) 2009-10-02 2016-06-21 Johnson & Johnson Consumer Inc. High-clarity aqueous concentrates of 4-hexylresorcinol
WO2016113357A1 (fr) * 2015-01-14 2016-07-21 Université D'aix-Marseille Inhibiteurs de protéasome pour le traitement d'un trouble lié à une accumulation de protéine anormale non dégradée ou d'un cancer
CN106974915A (zh) * 2017-04-01 2017-07-25 温州医科大学附属第二医院、温州医科大学附属育英儿童医院 一种雷公藤红素在制备治疗及延缓椎间盘退行性病变药物中的应用
US10307352B2 (en) 2012-09-24 2019-06-04 Johnson & Johnson Consumer Inc. Low oil compositions comprising a 4-substituted resorcinol and a high carbon chain ester
WO2022197773A1 (fr) * 2021-03-16 2022-09-22 Rush University Medical Center Méthodes de traitement de troubles neurodégénératifs au moyen d'un peptide de domaine de liaison (nbd) à un modificateur essentiel (nemo) de nf-kappab intranasal

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EP3302468A4 (fr) 2015-06-01 2019-02-06 The Scripps Research Institute Analogues à petites molécules du peptide de liaison nemo
US9943566B2 (en) 2016-03-16 2018-04-17 Geoffrey Brooks Consultants, Llc NF-κB inhibitor composition for skin health
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US9370474B2 (en) 2009-10-02 2016-06-21 Johnson & Johnson Consumer Inc. High-clarity aqueous concentrates of 4-hexylresorcinol
EP2316413A3 (fr) * 2009-10-02 2013-05-22 Johnson and Johnson Consumer Companies, Inc. Compositions comprenant un inhibiteur de NFkB et un promoteur du collagène non rétinoïde
US9289361B2 (en) 2009-10-02 2016-03-22 Johnson & Johnson Consumer Inc. Compositions comprising an NFκB-inhibitor and a non-retinoid collagen promoter
US9375395B2 (en) 2009-10-02 2016-06-28 Johnson & Johnson Consumer Inc. Compositions comprising an NFκB-inhibitor and a tropoelastin promoter
US8906432B2 (en) 2009-10-02 2014-12-09 Johnson & Johnson Consumer Companies, Inc. Compositions comprising an NFκB-inhibitor and a non-retinoid collagen promoter
RU2572505C2 (ru) * 2009-10-02 2016-01-10 Джонсон Энд Джонсон Конзьюмер Компаниз, Инк. КОМПОЗИЦИИ, ВКЛЮЧАЮЩИЕ ИНГИБИТОР NFкВ И СТИМУЛЯТОР ОБРАЗОВАНИЯ ТРОПОЭЛАСТИНА
EP2319488A3 (fr) * 2009-10-02 2013-06-12 Johnson & Johnson Consumer Companies, Inc. Compositions comprenant un inhibiteur de NFkB et un promoteur de la tropoélastine
RU2501552C2 (ru) * 2010-08-31 2013-12-20 Федеральное государственное бюджетное учреждение науки Институт биологии Коми научного центра Уральского отделения Российской академии наук Средство для увеличения продолжительности жизни и способ его применения
US8895628B2 (en) 2010-10-25 2014-11-25 Johnson & Johnson Consumer Companies, Inc. Compositions comprising a retinoid and an NFkB-inhibitor and their methods of use
US8877820B2 (en) 2010-10-25 2014-11-04 Johnson & Johnson Consumer Companies, Inc. Compositions comprising a retinoid and an NFkB-inhibitor and their methods of use
EP2572701A3 (fr) * 2010-10-25 2013-10-16 Johnson & Johnson Consumer Companies Inc. Compositions comprenant un rétinoïde et inhibiteur de NFkB et leurs procédés d'utilisation
US10307352B2 (en) 2012-09-24 2019-06-04 Johnson & Johnson Consumer Inc. Low oil compositions comprising a 4-substituted resorcinol and a high carbon chain ester
WO2016113357A1 (fr) * 2015-01-14 2016-07-21 Université D'aix-Marseille Inhibiteurs de protéasome pour le traitement d'un trouble lié à une accumulation de protéine anormale non dégradée ou d'un cancer
AU2016208001B2 (en) * 2015-01-14 2021-05-13 Assistance Publique Hopitaux De Marseille Proteasome inhibitors for treating a disorder related to an accumulation of non-degraded abnormal protein or a cancer
CN106974915A (zh) * 2017-04-01 2017-07-25 温州医科大学附属第二医院、温州医科大学附属育英儿童医院 一种雷公藤红素在制备治疗及延缓椎间盘退行性病变药物中的应用
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